Seqanswers Leaderboard Ad

Collapse

Announcement

Collapse
No announcement yet.
X
 
  • Filter
  • Time
  • Show
Clear All
new posts

  • RNA seq Library Purification

    Dear All!

    Hello, I am working on RNA sequencing NextSeq500 TrueSeq mRNA LT protocol. I have residual adapters after AmPure purification in 2/8 samples. Should I gel extract my these two or all samples or purify with AmPure beads again?

    I am wondering, if I might loose sample with AmPure purification. I used .85X beads ratio already.


    Any Recommendations? (Adapter dimer starts at 110bp and my library at 200bp)

    Regards
    Sahil Gupta
    Attached Files

  • #2
    Any purification method will cause loss of DNA with beads having relatively less loss than gel extraction. 0.85x bead should have cleaned the dimers if the protocol was followed without modification using calibrated pipettes. Two options:
    1- clean all samples if they are from the same experiment to reduce introducing another variable
    2- sequence samples with dimers deeper to compensate for the dimer reads that will be cleaned from data

    Comment


    • #3
      Thank you! It works with .8X beads purification!!!

      Comment

      Latest Articles

      Collapse

      • seqadmin
        Strategies for Sequencing Challenging Samples
        by seqadmin


        Despite advancements in sequencing platforms and related sample preparation technologies, certain sample types continue to present significant challenges that can compromise sequencing results. Pedro Echave, Senior Manager of the Global Business Segment at Revvity, explained that the success of a sequencing experiment ultimately depends on the amount and integrity of the nucleic acid template (RNA or DNA) obtained from a sample. “The better the quality of the nucleic acid isolated...
        03-22-2024, 06:39 AM
      • seqadmin
        Techniques and Challenges in Conservation Genomics
        by seqadmin



        The field of conservation genomics centers on applying genomics technologies in support of conservation efforts and the preservation of biodiversity. This article features interviews with two researchers who showcase their innovative work and highlight the current state and future of conservation genomics.

        Avian Conservation
        Matthew DeSaix, a recent doctoral graduate from Kristen Ruegg’s lab at The University of Colorado, shared that most of his research...
        03-08-2024, 10:41 AM

      ad_right_rmr

      Collapse

      News

      Collapse

      Topics Statistics Last Post
      Started by seqadmin, Yesterday, 06:37 PM
      0 responses
      11 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, Yesterday, 06:07 PM
      0 responses
      10 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 03-22-2024, 10:03 AM
      0 responses
      51 views
      0 likes
      Last Post seqadmin  
      Started by seqadmin, 03-21-2024, 07:32 AM
      0 responses
      67 views
      0 likes
      Last Post seqadmin  
      Working...
      X